Early molecular events in the assembly of the focal adhesion-stress fiber complex during fibroblast spreading

Cell Motil Cytoskeleton. 2004 Jul;58(3):143-59. doi: 10.1002/cm.20005.


Cell adhesion to the extracellular matrix triggers the formation of integrin-mediated contact and reorganization of the actin cytoskeleton. Examination of nascent adhesions, formed during early stages of fibroblast spreading, reveals a variety of forms of actin-associated matrix adhesions. These include: (1). small ( approximately 1 microm), dot-like, integrin-, vinculin-, paxillin-, and phosphotyrosine-rich structures, with an F-actin core, broadly distributed over the ventral surfaces of the cells; (2). integrin-, vinculin-, and paxillin-containing "doublets" interconnected by short actin bundles; (3). arrays of actin-vinculin complexes. Such structures were formed by freshly plated cells, as well as by cells recovering from latrunculin treatment. Time-lapse video microscopy of such cells, expressing GFP-actin, indicated that long actin cables are formed by an end-to-end lining-up and apparent fusion of short actin bundles. All these structures were prominent during cell spreading, and persisted for up to 30-60 min after plating. Upon longer incubation, they were gradually replaced by stress fibers, associated with focal adhesions at the cell periphery. Direct examination of paxillin and actin reorganization in live cells revealed alignment of paxillin doublets, forming long and highly dynamic actin bundles, undergoing translocation, shortening, splitting, and convergence. The mechanisms underlying the assembly and reorganization of actin-associated focal adhesions and the involvement of mechanical forces in regulating their dynamic properties are discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / metabolism
  • Actins / metabolism*
  • Animals
  • Bridged Bicyclo Compounds, Heterocyclic / pharmacology
  • Cell Adhesion / physiology
  • Cell Movement / physiology*
  • Cells, Cultured
  • Cytoskeletal Proteins / metabolism
  • Cytoskeleton / metabolism
  • Cytoskeleton / ultrastructure
  • Extracellular Matrix / metabolism
  • Fibroblasts / physiology*
  • Fibroblasts / ultrastructure
  • Focal Adhesions / physiology*
  • Focal Adhesions / ultrastructure
  • Humans
  • Integrin alphaVbeta3 / metabolism
  • Integrins / metabolism
  • Male
  • Microscopy, Electron, Scanning
  • Microscopy, Video
  • Paxillin
  • Phosphoproteins / metabolism
  • Phosphotyrosine / metabolism
  • Rats
  • Stress Fibers / physiology*
  • Stress Fibers / ultrastructure
  • Stress, Mechanical
  • Thiazoles / pharmacology
  • Thiazolidines
  • Time Factors
  • Vinculin / metabolism


  • Actins
  • Bridged Bicyclo Compounds, Heterocyclic
  • Cytoskeletal Proteins
  • Integrin alphaVbeta3
  • Integrins
  • PXN protein, human
  • Paxillin
  • Phosphoproteins
  • Pxn protein, rat
  • Thiazoles
  • Thiazolidines
  • Vinculin
  • Phosphotyrosine
  • latrunculin A